One-Bead-One-Peptide Library to Purify Crotalus durissus terrificus Phosholipase A2

MARTÍNEZ CERON, M. C.; SAAVEDRA, S. L.; LUCÍA ÁVILA; SILVANA L. GIUDICESSI; FERNANDO ALBERICIO; SILVIA A. CAMPERI; OSVALDO CASCONE

Orlando

Simposio; 24th Internattional America Peptide Symposium; 2015

American Peptide Society

Crotalus durissus terrificus (Cdt), is the only species of that genus in Argentina. Crotoxin represents more than 50% of the dry weight of its poison. This protein has two subunits: A (phospholipase A2, PLA2), and B (crotapotin). Poison profiles differ according to geographical regions. The aim of this work was to purify Argentinean Cdt PLA2 to analyze their characteristics and possible uses as antiviral activity against tropical diseases. Short peptides are stable and resistant to proteases and can be produced in high quantities and purity. In this work, a one-bead-one-peptide library (XXXXXGGAGG; X= A, E, F, H, L, N, P, R, S, T, V, Y) was synthesized using Divide-Couple-Recombine method (DCR). Screening was done using pure PLA2 labeled with NHS-Biotin. Those peptides with affinity for the protein were revealed using Strepatavidin-Peroxidase and Alpha-Chloronaphtol/H2O2. They were isolated to release the peptides. Peptides were analyzed by MALDI-TOF MSMS. Fifty peptides were sequenced and their amino acids moieties were studied. Two of them were selected to be immobilized in Sepharose. Both peptides were capable to purify PLA2 in one step with high yield.